PeHDZ23987 activated PeAAP29123 by binding to its promoter. (IMAGE)

Nanjing Agricultural University The Academy of Science

PeHDZ23987 activated PeAAP29123 by binding to its promoter.

Caption

PeHDZ23987 activated PeAAP29123 by binding to its promoter. (A) The distribution diagram of TF binding sites of proPeAAP29123. (B) Y1H assays were used to identify PeHDZ23987 binding to the promoter of PeAAP29123. (C) Dual-luciferase complementation imaging assays used to show that PeHDZ23987 activated the expression of PeAAP29123. (D) The firefly luciferase (LUC)/Renilla luciferase (REN) (LUC/REN) activity ratio was used to verify that PeHDZ23987 activated PeAAP29123. (E, G) PeHDZ23987 activating the transcription of HD-motifs of PeAAP29123 in tobacco leaves by dual-luciferase assays. pMotif1, pMotif2, pMutant1, and pMutant2 indicated pPeAAP29123-Motif1::LUC, pPeAAP29123-Motif2::LUC, pPeAAP29123-Mutant2::LUC, and pPeAAP29123-Mutant1::LUC, respectively. Representative photographs were taken using a chemiluminescence imaging system. (F, H) LUC/REN activity detection to verify PeHDZ23987 activating the transcription of HD-motifs of PeAAP29123. Data are shown as mean ± standard deviation (n ≥ 3). The asterisk indicates a significant difference (*P < 0.05, **P < 0.01). (I–J) PeHDZ23987 binds to two motifs (HDZ-motif1 5'-CAATCAAG-3′ and HDZ-motif2 5'-CAATTATT-3′) of the PeAAP29123 promoter by EMSA. Unlabelled probes were used as competitors. The 60× represent the concentrations of the competitor. Mut represents a mutated probe in which the motif is replaced by 5'-ACGGCTCC-3′.

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Horticulture Research

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